Piperidine-2, 6-dione derivatives and their use as tumor necrosis factor inhibitors

ABSTRACT

This invention is directed to derivatives of piperidine-2,6-dione, or their organic or inorganic salts thereof, a methods of synthesis of these derivatives, and their application as active pharmaceutical ingredient as inhibitors of TNFα releasing in cells, the derivative of piperidine-2,6-dione being of the general formula (I): 
                         
wherein n represents 1, 2, 3, 4, 5 or 6; R 1  represents from one to four of the same or different substituents selected from F, Cl, Br, C 1-4  alkyl, OH, OC 1-4  alkyl, NO 2 , NHC(O)C 1-4  alkyl, NH 2 , NH(C 1-4  alkyl), N(C 1-4  alkyl) 2 ; R 2  represents OR 3 , NR 3 R 4 , N(R 3 )COR 4 , O 2 CR 5 ; R 3  and R 4  represent independently and at each occurrence H or C 1-4  alkyl; R 5  represents CHR 6 NR 7 R 8 , CHR 6 NR 9 C(O)CHR 10 NR 7 R 8 , a heterocycle W or CHR 6 NR 9 C(O)W; R 6 , R 9 , R 10  represent independently and at each occurrence H, or C 1-4  alkyl; R 7  and R 8  represent independently and at each occurrence H, C 1-4  alkyl, or R 7  and R 8  taken together represent 1,3-propylene, 1,4-butylene, 1,5-pentylene, or 1,6-hexylene; W represents four-membered, five-membered, six-membered, seven-membered, or eight-membered saturated or unsaturated heterocycle.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International Patent ApplicationNo. PCT/CN2005/001467 with an international filing date of Sep. 13,2005, designating the United States, now pending, and further claimspriority benefits to Chinese Patent Application No 200510013292.3, filedApr. 7, 2005. The contents of the aforementioned specifications areincorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention concerns piperidine-2,6-dione derivatives which areactive as inhibitors of tumor necrosis factor released by cells, thepreparation of these derivatives, as well as their application aspharmaceutically-active ingredients.

2. Description of the Related Art

Tumor necrosis factor α (TNFα) is a cytokine, mainly produced bymacrophages, which causes inflammation, fever, cardiovasculardysfunction, hemorrhage and a series of acute reactions similar to acuteinfection and shock when it is applied in humans and animals. Excessiveor uncontrolled TNFα in animals or humans often indicates one of thefollowing diseases:

-   -   1) Endotoxaemia and/or toxic shock syndrome [Tracey et al.,        Nature 330, 662-4 1987; Hinshaw et al., Circ Shock 30, 279-92        (1990)];    -   2) Cachexia [Dezube et al., Laucet, 335(8690), 662 (1990)]; or    -   3) Adult Respiratory Distress Syndrome (ARDS) [Millar et al.,        Laucet 2(8665), 712-714 (1989)].

TNFα also plays an important role in bone resorption diseases includingarthritis [Betolinni et al., Nature 319, 516-8 (1986)]. TNFα maystimulate bone resorption by stimulating the formation and activation ofosteoclast and resist the formation of bone, which was shown both by invitro and in vivo experiments.

At the present, a disease which is most commonly linked to TNFα releasedby tumor and host tissue is hypercalcemia, which is closely related tomalignant tumors [Calci. Tissue Int. (US) 46(Suppl.), S3-10 (1990)]. Theimmune response is closely related to an increased concentration of TNFαin serum of the patient after bone marrow transplantation [Holler etal., Blood, 75(4), 1011-1016 (1990)].

Fatal hyperacute neurogenic syndrome brainstem-type malaria, which isthe most dangerous type of malaria, is also linked to high levels ofTNFα in blood. When this kind of malaria occurs, the levels of TNFα inserum is directly related to the disease, which often occurs during anacute attack of malaria in patients [Grau et al., N. Engl. J. Med.320(24), 1586-91 (1989)].

TNFα plays an important role in chronic pneumonia as well. The storageof silicon-containing particles can cause silicosis. Silicosis is apulmonary fibrosis, which causes progressive respiratory failure. In ananimal pathological model, TNFα antibody can fully block the progress ofmice lung fibrosis caused by silica dust [Pignet et al., Nature,344:245-7 (1990)]. It was also proved that TNFα levels are abnormallyhigh in serum of animals with pulmonary fibrosis caused by silica dustor asbestos dust in animal experiments [Bissonnette et al., Inflammation13(3), 329-339 (1989)]. Pathological research reveals that TNFα levelsin the lungs of Pneumal Sarcoidosis patients is much higher than that ofordinary people [Baughman et al., J. Lab. Clin. Med. 115(1), 36-42(1990)]. It follows that TNFα inhibitor should have a great significancein the treatment of chronic pulmonary disease and lung injury.

The reason for inflammation occurring in the body of patient havingreperfusion injury may be abnormal levels of TNFα. TNFα is regarded asthe chief cause inducing tissue injury caused by ischemia [Uadder etal., PNAS 87, 2643-6 (1990)].

Besides, it has been shown that TNFα may start retroviral replicationincluding that of HIV-1 [Duh et al., Proc. Nat. Acad. Sci., 86, 5974-8(1989)]. T-cells need to be activated before HIV infects them. Once theactivated T-cells are infected by virus (HIV), those T-cells must be inan activated state so that HIV virus genes are able to be expressedand/or replicated successfully. Cytokines, especially TNFα, play animportant role in the process of HIV protein expression or viralreplication controlled by T-cells. So, inhibition of TNFα formation canin turn inhibit HIV replication in T-cells [Poll et al., Proc. Nat.Acad. Sci., 87, 782-5 (1990); Monto et al., Blood 79, 2670 (1990); Pollet al., AIDS Res. Human Retrovirus, 191-197 (1992)].

cAMP can control many functions of cells, such as inflammation response,including asthma, and inflammation [Lome and Cheng, Drugs of the futune,17(9), 799-807, 1992]. When inflammation occurs, increased cAMPconcentration in white cells inhibits white cell activation and thenreleases inflammation regulatory factors including TNFα so as toexacerbate inflammation in patients. Consequently, the inhibition ofTNFα release can alleviate inflammation diseases including asthma.

Several doctors, including Yu Yanyan, have found that TNFα plays animportant role in the process of liver necrosis in viral hepatitispatients. [Yu Yanyan etc., Chinese Journal of Internal Medicine 1996,35:28-31]. This shows that TNFα inhibitors may play a great role in thetreatment of chronic hepatic disease and liver injury.

Several researchers, including Li Yingxu, have found that levels oftumor necrosis factors are significantly increased on synthesis andsecretion of human monocyte in patients with chronic hepatic disease andother cell factor secretions are induced (for example, II-1β, II-6 andII-8). They are both involved in hepatocellular injury process [Journalof Qiqihar Medical Colleg, 22(10):1119-1120, 2001]. Their results are inaccordance with the conclusions of Yoshioka etc. [Hepatology, 1989,10:769-777] and Wang Xin etc. [Chinese Journal of Infectious Diseases,1997, 15(2):85-88]. It has also been found that thalidomide, the smallmolecular inhibitor of TNFα, is able to inhibit TNFα secreted by humanmonocyte in hepatitis patients, which lays a foundation of molecularpathology for TNFα inhibitor applied on hepatitis, cirrhosis and livercancer therapy.

TNFα induces certain inflammation responses, such as aggregation andadhesion of inflammatory cells, increased dilatation and permeability ofmicro-vessels, fever, increased neutrophil in circulation, andhemodynamic changes, and further causes kidney cell injury bystimulating synthesis and release of inflammation cytokine [Abboud H. E.Kidney Int. 1993; 43:252-267], expression of cell adhesion molecule[Egido J. et al., Kidney Int. 1993; 43(suppl 39):59-64], synthesis andrelease of prostaglandin G₂ (PGE₂) and platelet-activating factor(PAF)[Gammusi G. et al., Kidney Int., 43(suppl 39):32-36]. It has beenshown that TNFα plays an important role in the development of nephritis.

TNFα regulates the differentiation of B lymphocytes and reinforces thecytotoxicity of natural killer cells (NK), so as to participate in theregulation of immunological function by the activation of hyperplasia ofmacrophages and immunologically stimulating T-lymphocytes.

Therefore, it is an effective strategy to decrease TNFα levels and/orincrease cAMP levels so as to cure many inflammatory, infectious,immunological or malignant tumor diseases, including but not limited toseptic shock, endotoxic shock, hemodynamic shock, septic syndrom, postischemic reperfusion injury, malaria, mycobacterial infection,meningitis, psoriasis, congestive heart failure, fibrotic disease,cachexia, transplant immune rejection, cancer, autoimmune disease,opportunistic infection in AIDS, rheumatoid arthritis (RA), hepatitis,nephritis, rheumatoid spondylitis, and so on. Accordingly, research anddevelopment on small molecular TNFα inhibitors with low toxicity andhigh efficiency has a great public significance and economic value.

SUMMARY OF THE INVENTION

In one aspect, this invention is directed to derivatives ofpiperidine-2,6-dione, their organic or inorganic salts thereof, methodsto synthesize these derivatives, and their application aspharmaceutically-active ingredients as inhibitors of TNFα releasing incells, the derivatives of piperidine-2,6-dione being of the generalformula (I):

wherein n represents 1, 2, 3, 4, 5 or 6; R¹ represents from one to fourof the same or different substituents selected from F, Cl, Br, C₁₋₄alkyl, OH, OC₁₋₄ alkyl, NO₂, NHC(O)C₁₋₄ alkyl, NH₂, NH(C₁₋₄ alkyl),N(C₁₋₄ alkyl)₂; R² represents OR³, NR³R⁴, N(R³)COR⁴, O₂CR⁵; R³ and R⁴represent independently and at each occurrence H or C₁₋₄ alkyl; R⁵represents CHR⁶NR⁷R⁸, CHR⁶NR⁹C(O)CHR¹⁰NR⁷R⁸, a heterocycle W orCHR⁶NR⁹C(O)W; R⁶, R⁹, R¹⁰ represent independently and at each occurrenceH, or C₁₋₄ alkyl; R⁷ and R⁸ represent independently and at eachoccurrence H, C₁₋₄ alkyl, or R⁷ and R⁸ taken together represent1,3-propylene, 1,4-butylene, 1,5-pentylene, or 1,6-hexylene; Wrepresents a four-membered, a five-membered, a six-membered, aseven-membered, or an eight-membered saturated or unsaturatedheterocycle.

When W represents a heterocycle, it includes a four-membered, afive-membered, a six-membered, a seven-membered or an eight-memberedsaturated, or unsaturated heterocycle or aromatic heterocycle, bearingone or multiple heteroatoms such as nitrogen atom, oxygen atom or sulfuratom, and particularly, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl,3-pyrimidinyl, 4-pyrimidinyl, or heterocycles of formula (II), (III),(IV), or (V), in which X represents O, S, or NR¹²; Y represents1,2-ethylene, 1,3-propylene, 1,4-butylene, 1,5-pentylene, 1,6-hexylene,—CH₂OCH₂—, —CH₂SCH₂— or —CH₂NR¹²CH₂—; and R¹¹ and R¹² representindependently and at each occurrence H, or C₁₋₄alkyl. R³, R⁴, R⁶, R⁹,R¹⁰, R¹¹, or R¹² may be substituted by substituents

such as OH, COOH, C(O)NH₂, NHC(O)R¹³, NH₂, NHR¹⁴, NR¹⁵R¹⁶, NHC(O)NH₂,NHC(NH)NH₂, OR¹⁷, SR¹⁸, phenyl or substituted phenyl, etc, when R³, R⁴,R⁶, R⁹, R¹⁰, R¹¹, R¹² represent C₁₋₄ alkyl, including straight chain orbranched chain alkyl, wherein R¹³R¹⁴, R¹⁵, R¹⁶, R¹⁷ and R¹⁸ representindependently and at each occurrence H, or C₁₋₄ alkyl.

R⁷ and R⁸ may be substituted by substituents such as OH, COOH, C(O)NH₂,NHC(O)R¹³, NH₂, NHR¹⁴, NR¹⁵R¹⁶, NHC(O)NH₂, NHC(NH)NH₂, OR¹⁷, SR¹⁸,phenyl, or substituted phenyl, etc, when R⁷ and R⁸ independentlyrepresent C₁₋₄ alkyl, including straight chain or branched chain alkyl,wherein R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, and R¹⁸ represent independently and ateach occurrence H, or C₁₋₄ alkyl.

R⁷ and R⁸ may be substituted by substituents such as OH, COOH, C(O)NH₂,NHC(O)R¹³, NH₂, NHR¹⁴, NR¹⁵R¹⁶, NHC(O)NH₂, NHC(NH)NH₂, OR¹⁷, SR¹⁸,phenyl or substituted phenyl, etc., when R⁷ and R⁸ taken togetherrepresent 1,3-propylene, 1,4-butylene, 1,5-pentylene, or 1,6-hexylene,wherein R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, and R¹⁸ represent independently and ateach occurrence H, or C₁₋₄ alkyl.

When W represents a heterocycle, it includes four-membered,five-membered, six-membered, seven-membered or eight-membered saturated,or unsaturated heterocycles or aromatic heterocycles including one ormultiple heteroatoms, such as nitrogen atom, oxygen atom or sulfur atom,which can be substituted by OH, COOH, C(O)NH₂, NHC(O)R¹³, NH₂, NHR¹⁴,NR¹⁵R¹⁶, NHC(O)NH₂, NHC(NH)NH₂, OR¹⁷, SR¹⁸ OR¹⁹, wherein R¹³, R¹⁴, R¹⁵,R¹⁶, R¹⁷, R¹⁸, R¹⁹ represent independently and at each occurrence H, orC₁₋₄ alkyl.

The compounds of formula (I) appropriate for medical use include thosecompounds wherein n represents an integer from one to six, andparticularly those compounds, wherein n represents one, two or three.

The compounds of formula (I) appropriate for medical use include thosecompounds wherein R¹ represents from one to four of same or differentsubstituents selected from: H, F, Cl, Br, CH₃, CH₂CH₃, OH, OCH₃,OCH₂CH₃, NH₂, NHCH₃, NHCH₂CH₃, N(CH₃)₂; and particularly the compoundswherein R¹ represents H, 3-F, 4-F, 3-NH₂, 4-NH₂, or 3,4,5,6-tetrafluoro.

The compounds of formula (I) appropriate for medical use include thosecompounds wherein R⁵ represents CHR⁶NR⁸R⁷; R⁶ represents H, CH₃, CH₂CH₃,CH₂CH₂CH₃, CH(CH₃)₂, CH₂CH₂CH₂CH₃, CH₂CH(CH₃)₂, or CH(CH₃)CH₂CH₃; R⁷ andR⁸ represent independently and at each occurrence H, CH₃, CH₂CH₃,CH₂CH₂CH₃, CH(CH₃)₂, CH₂CH₂CH₂CH₃, CH₂CH(CH₃)₂, or CH(CH₃)CH₂CH₃; R⁷ andR⁸ taken together represent 1,4-butylene, 1,5-pentylene; andparticularly those compounds wherein R⁶ represents H, CH₃, or CH(CH₃)₂;R⁷ and R⁸ represent independently and at each occurrence H, CH₃, CH₂CH₃;or R⁷ and R⁸ taken together represent 1,4-butylene, or 1,5-pentylene.

When R⁵ represents CHR⁶NR⁹C(O)CHR¹⁰NR⁷R⁸, the compounds of formula (I)appropriate for medical use include those in which R⁶ and R¹⁰independently and at each occurrence represent H, CH₃, CH(CH₃)₂,CH₂CH(CH₃)₂, or CH(CH₃)CH₂CH₃; R⁹ represents H, CH₃, CH₂CH₃, CH₂CH₂CH₃,CH(CH₃)₂, R⁸ and R⁷ each independently and at each occurrence representH, CH₃, CH₂CH₃, CH₂CH₂CH₃, CH(CH₃)₂, or R⁸ and R⁷ taken togetherrepresent 1,4-butylene, or 1,5-pentylene; and particularly compounds offormula (I) appropriate for medical use include those derivatives ofpiperidine-2,6-dione in which R⁶ and R¹⁰ independently and at eachoccurrence represent H, CH₃ or CH(CH₃)₂, R⁹ represents H, CH₃, CH₂CH₃;and R⁸ and R⁷ independently and at each occurrence represent H, CH₃,CH₂CH₃ or R⁸ and R⁷ taken together represent 1,4-butylene or1,5-pentylene.

The compounds of formula (I) appropriate for medical use include thosecompounds wherein R⁵ represents W, and W represents 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl,2-pyrrolidinyl, 2-(N-methyl)pyrrolidinyl, 2-(N-ethyl)pyrrolidinyl,2-(N-propyl)pyrrolidinyl, or 2-(N-isopropyl)pyrrolidinyl. Among them,those particularly appropriate for medical use include those compoundsin which W represents 3-pyridyl, 2-pyrrolidinyl,2-(N-methyl)pyrrolidinyl, or 2-(N-ethyl)pyrrolidinyl.

The compounds of formula (I) appropriate for medical use include thosederivatives of piperidine-2,6-dione wherein R⁵ represents CHR⁶NR⁹C(O)W.

Among the compounds of formula (I) appropriate for medical use whereinR⁵ represents CHR⁶NR⁹C(O)W are included derivatives ofpiperidine-2,6-dione wherein R⁶ and R⁹ independently and at eachoccurrence represent H, CH₃, CH₂CH₃, CH₂CH₂CH₃ or CH(CH₃)₂, and Wrepresents 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl,4-pyrimidinyl, 5-pyrimidinyl, 2-pyrrolidinyl, 2-(N-methyl)pyrrolidinyl,2-(N-ethyl)pyrrolidinyl, 2-(N-propyl)pyrrolidinyl, or2-(N-isopropyl)pyrrolidinyl. Those compounds particularly appropriatefor medicinal purposes include compounds of formula (I) wherein R⁶represents H, CH₃ or CH(CH₃)₂, R⁹ represents H, CH₃, CH₂CH₃; and Wrepresents 3-pyridyl, 2-pyrrolidinyl, 4-pyridyl,2-(N-methyl)-pyrrolidinyl, or 2-(N-ethyl)pyrrolidinyl.

Particular derivatives of piperidine-2,6-dione appropriate for medicaluse as active ingredient include but are not limited to the followingcompounds:

-   1)    4-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   2)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   3)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   4)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   5)    5-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione-   6)    2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;-   7)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;-   8)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   9)    2-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   10)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(dimethylamino)acetate;-   11)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(diethylamino)acetate;-   12)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(1-piperidyl)acetate;-   13)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-aminoacetate;-   14)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-aminopropanoate;-   15)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-amino-3-methylbutanoate;-   16)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidinecarboxylate;-   17)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(2-diethylaminoacetamido)propanoate;-   18)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(2-diethylaminoacetamido)-3-methylbutanoate;-   19)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-amino-3-phenylpropanoate;-   20)    2-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(diethylamino)acetate;-   21)    4-amino-2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   22)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    nicotinate;-   23)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    isonicotinate;-   24)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-ethyl-(S)-2-pyrrolidinecarboxylate;-   25)    4-amino-2-(2,6-dioxo-1-(2-propoxyethyl)piperidin-3-yl)isoindoline-1,3-dione;-   26)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-5-nitroisoindoline-1,3-dione;-   27)    4,5,6,7-tetrafluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   28)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    nicotinate;-   29)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    acetate;-   30)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    isonicotinate;-   31)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-ethyl-(S)-2-pyrrolidinecarboxylate;-   32)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidinecarboxylate;-   33)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(diethylamino)acetate;-   34)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(dimethylamino)acetate;-   35)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(ethylamino)acetate;-   36)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(methylamino)acetate;-   37)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-aminoacetate;-   38)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(1-piperidyl)acetate;-   39)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(diethylamino)propanoate;-   40)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(dimethylamino)propanoate;-   41)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-ethylaminopropanoate;-   42)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-methylaminopropanoate;-   43)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-aminopropanoate;-   44)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(pyrrolidin-1-yl)propanoate;-   45)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(piperidin-1-yl)propanoate;-   46)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(pyrrolidin-1-yl)acetate;-   47)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-diethylamino-3-methylbutanoate;-   48)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-dimethylamino-3-methylbutanoate;-   49)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-ethylamino-3-methylbutanoate;-   50)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-methylamino-3-methylbutanoate;-   51)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-amino-3-methylbutanoate;-   52)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidin-1-yl-3-methylbutanoate;-   53)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-piperidin-1-yl-3-methylbutanoate;-   54)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    nicotinate;-   55)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    acetate;-   56)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    isonicotinate;-   57)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-ethyl-(S)-2-pyrrolidinecarboxylate;-   58)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidinecarboxylate;-   59)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(diethylamino)acetate;-   60)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(dimethylamino)acetate;-   61)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(ethylamino)acetate;-   62)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(methylamino)acetate;-   63)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-aminoacetate;-   64)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(1-piperidyl)acetate;-   65)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(diethylamino)propanoate;-   66)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(dimethylamino)propanoate;-   67)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(ethylamino)propanoate;-   68)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(methylamino)propanoate;-   69)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-aminopropanoate;-   70)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(pyrrolidin-1-yl)propanoate;-   71)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(piperidin-1-yl)propanoate;-   72)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(pyrrolidin-1-yl)acetate-   73)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-diethylamino-3-methylbutanoate;-   74)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-dimethylamino-3-methylbutanoate;-   75)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-ethylamino-3-methylbutanoate;-   76)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-methylamino-3-methylbutanoate;-   77)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-amino-3-methylbutanoate;-   78)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidin-1-yl-3-methylbutanoate;-   79)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-piperidin-1-yl-3-methylbutanoate;-   80)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    nicotinate;-   81)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    acetate;-   82)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    isonicotinate;-   83)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-ethyl-(S)-2-pyrrolidinecarboxylate;-   84)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidinecarboxylate;-   85)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(diethylamino)acetate;-   86)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(dimethylamino)acetate;-   87)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(ethylamino)acetate;-   88)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(methylamino)acetate;-   89)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-aminoacetate;-   90)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(1-piperidyl)acetate;-   91)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(diethylamino)propanoate;-   92)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(dimethylamino)propanoate;-   93)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-ethylaminopropanoate;-   94)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-methylaminopropanoate;-   95)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-aminopropanoate;-   96)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(pyrrolidin-1-yl)propanoate;-   97)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-(piperidin-1-yl)propanoate;-   98)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(pyrrolidin-1-yl)acetate;-   99)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-diethylamino-3-methylbutanoate;-   100)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-dimethylamino-3-methylbutanoate;-   101)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-ethylamino-3-methylbutanoate;-   102)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-methylamino-3-methylbutanoate;-   103)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-amino-3-methylbutanoate;-   104)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidin-1-yl-3-methylbutanoate;-   105)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-piperidin-1-yl-3-methylbutanoate;-   106)    4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-methyl-(S)-2-pyrrolidinecarboxylate;-   107)    4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-methyl-(S)-2-pyrrolidinecarboxylate;-   108)    5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-methyl-(S)-2-pyrrolidinecarboxylate;-   109)    2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;-   110)    4-(dimethylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   111)    4-(methylamino)-2-(2,6-dioxo-1-(2-propoxyethyl)piperidin-3-yl)isoindoline-1,3-dione;-   112)    2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;-   113)    2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;-   114)    N-(2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-7-yl)acetamide;-   115)    4-(dimethylamino)-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   116)    4-(dimethylamino)-2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   117)    4-(dimethylamino)-2-(1-(3-hydroxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   118)    4-(dimethylamino)-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   119)    4-amino-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   120)    4-amino-2-(1-(3-hydroxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   121)    5-amino-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;-   122)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    nicotinate;-   123)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    acetate;-   124)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    isonicotinate;-   125)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    N-ethyl-(S)-2-pyrrolidinecarboxylate;-   126)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-pyrrolidinecarboxylate;-   127)    4-(methylamino-)2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(diethylamino)acetate;-   128)    4-(methylamino-)2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(dimethylamino)acetate;-   129)    4-(methylamino-)2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(ethylamino)acetate;-   130)    4-(methylamino-)2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(methylamino)acetate;-   131)    4-(methylamino-)2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-aminoacetate;-   132)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    2-(1-piperidyl)acetate;-   133)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-amino-3-methylbutanoate; and-   134)    4-(methylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione    (S)-2-aminopropanoate.

The compound of the invention shown in formula (I) and used as apharmaceutically active ingredient may be a single enantiomer (R or S)or racemate.

The compounds of this invention shown in formula (I) may be used aspharmaceutically-active ingredients in various forms, for example, freebases, inorganic acid salts, which includes hydrochloride, sulfate,nitrate, phosphate, and also organic salts, which includes sulfonate,acetate, formate, fumarate, maleate, citrate, tartrate, malate,benzoate, ascorbate, gluconate, lactate, succinate and trifluoroacetate.

In other aspects, this invention is directed at a method to prepare thecompounds of the general formula (I), by reacting compounds bearinggeneral formula (VI),

with Z—CH₂(CH₂)_(n)—R² wherein R¹ represents from one to four of thesame or different substituents selected from F, Cl, Br, NO₂, H, C₁₋₄alkyl, OR³, NR³R⁴; n represents 1, 2, 3, 4, 5, or 6; R² represents OR³,NR³R⁴, N(R³)COR⁴, O₂CR⁵; R³ and R⁴ represent independently and at eachoccurrence H, or C₁₋₄ alkyl; R⁵ represents CHR⁶NR⁷R⁸,CHR⁶NR⁹C(O)CHR¹⁰NR⁷R⁸, W, or CHR⁶NR⁹C(O)W; R⁶, R⁹ and R¹⁰ independentlyand at each occurrence represent H, C₁₋₄ alkyl; R⁷ and R⁸ independentlyand at each occurrence represent H, C₁₋₄ alkyl, or R⁷ and R⁸ takentogether represent 1,3-propylene, 1,4-butylene, 1,5-pentylene, or1,6-hexylene; W represents four-membered, five-membered, six-membered,seven-membered or eight-membered saturated or unsaturated heterocycles,particularly 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl,3-pyrimidinyl, 4-pyrimidinyl, or heterocycles of the formula (II),(III), (IV) or (V);

X represents O, S, or NR¹²; Y represents substituents such as1,2-ethylene, 1,3-propylene, 1,4-butylene, 1,5-pentylene, 1,6-hexylene,—CH₂OCH₂—, —CH₂SCH₂—, or —CH₂NR¹²CH₂—; and R¹¹ and R¹² eachindependently and at each occurrence represent H, or C₁₋₄ alkyl; Zrepresents Cl, Br.

In a method to prepare the compounds of the general formula (I), theratio of a compound shown in Formula (VI) to Z—CH₂(CH₂)_(n)—R² may bebetween 3:1 and 1:3.

The synthesis may be facilitated by an inorganic base, which includesbut is not limited to NaH, KH, CaH₂, K₂CO₃, Na₂CO₃, KHCO₃, NaHCO₃,Li₂CO₃, Cs₂CO₃, LiOH, KOH, NaOH, Ca (OH)₂, K₃PO₄, K₂HPO₄, or an organicbase. The proportion of base to substrate is from 50% (mole) to 300%(mole). The reactions are conducted in an organic solvent, such asdichloromethane, chloroform, acetone, butanone, dimethylformamide,dimethylsulfoxide, ethylene glycol dimethyl ether, tetrahydrofuran,pyridine or acetonitrile, and may be conducted under heterogeneousconditions, especially with a phase-transfer catalyst.

The compounds of formula (I) are indicated for and are useful in thetreatment and prevention of diseases which are associated with decreasedTNFα levels in patients, including, but not limited to inflammatory orinfectious diseases, diseases of the immune system, or malignant tumors.Particularly, these diseases include but are not limited to septicshock, endotoxic shock, hemodynamic shock, septic syndrom, post ischemicreperfusion injury, malaria, mycobacterial infection, meningitis,psoriasis, congestive heart failure, fibrotic disease, cachexia,transplant immune rejection, cancer, autoimmune disease, opportunisticinfection in AIDS, erythema nodosum leprosum, lupus erythematosus,intractable lupus erythematosus, Behcet syndrome, regional ileitis,myelodysplastic syndrome, rheumatoid arthritis (RA), hepatitis,nephritis, rheumatoid spondylitis, multiple myeloma, thyroma, renalcancer, prostate carcinoma, lymphoma, leukemia, and hepatoma.

Except for at least one kind of compounds of formula (I), thepharmaceutical compositions of the invention may comprise one or morecarrier materials, bulking agents, solvents, diluents, colorants, and/oradhesives. The selection of adjuvants and dosage is decided taking intoaccount the mode of administration, e.g., gastrointestinal, intravenous,abdominal, dermal, intramuscular, nasal, ocular, pulmonary, anal,vaginal, transdermal, etc.

The pharmaceutical compositions and pharmaceutically active compounds ofthe invention may be used in combination with other appropriatepharmaceutically active compounds and/or pharmaceutical compositions.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1-9 illustrate chemical structures of piperidine-2,6-dionederivatives according to the invention; and

FIGS. 10-11 illustrate chemical structures of intermediates in thesynthesis of piperidine-2,6-dione derivatives and chemical structures ofpiperidine-2,6-dione derivatives according to the invention.

DETAILED DESCRIPTION OF THE INVENTION

Specific Implementation Method

Pharmacological Research: Effects of Monocyte (PBMC) Stimulation by LPSon TNFα

Cytokine TNFα released by PBMCs in peripheral blood after lippolysaccharide (LPS) stimulation in vitro was studied. The followingsare research methods of cytokine TNFα released by PBMCs, which areinhibited by compounds of the invention.

PBMCs were obtained from blood of at least three volunteers afterheparin treatment, by the method of gradient extraction. PBMCs werecollected and washed with 1640 culture medium three times (10% calfserum, 2 mM L-glutamine, 100 mM mercaptoethanol, 50 μg/ml streptomycin,50 U/ml penicillin). The above PBMCs were placed onto a 24-well cellculture plate. The concentration was adjusted to 1×10⁶ cells/ml with1640 culture medium. Test compounds were dissolved in dimethylsulfoxideat an appropriate concentration. The resultant solution was added to theabove cell culture medium and the culture plate was placed in a CO₂incubator (5% CO₂, 90% humidity) for 1 hour. Then, LPS (Sigma) was addedto adjust the concentration to 0.1 μg/ml (except for contrast). After 20hrs of incubation, the content of TNFα in supernatant of the above PBMCculture medium was determined by ELISA kit (America Genzyme Co) usingstandard method. The measured value of the control (no active compound),and the measured value of the tested compounds was used to calculate theTNFα inhibition rate. The concentration of compounds giving a 50% TNFαinhibition (IC₅₀ value) was calculated by nonlinear regression analysis.Each concentration was determined twice and an average value wasobtained. Results are illustrated in Table 1.

TABLE 1 Inhibit LPS to stimulate monocytes to inhibit TNFα activityCompound Concentration (μM) Inhibition Degree (%) EC₅₀ (μM) Thalidomide100 22 183 of example 1 3.0 70 of example 9 3.0 20 of example 22 3.0 18of example 24 3.0 28 of example 26 3.0 95 0.25 of example 27 3.0 92 0.3of example 28 3.0 78 of example 29 3.0 64 of example 30 3.0 58 ofexample 31 3.0 62

EXAMPLES

Abbreviations

-   DCC: dicyclohexylcarbodiimide; DCM: dichloromethane; TFA:    trifluoroacetic acid;-   CDCl₃: deuterochloroform; HCl: hydrogen chloride; DMAP:    4-(N,N-dimethylamino)pyridine; TEA: triethylamine.

Example 12-(1-(2-Hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

2-(2,6-Dioxopiperidin-3-yl)isoindoline-1,3-dione (2.5 g) was dissolvedin DMF (dry, 60 mL). 95% NaH (0.24 g) was added. The mixture was stirredfor 30 minutes at room temperature. Then, chloroethanol (0.68 mL) wasadded, and the mixture was stirred over night at room temperature. Thereaction mixture was diluted with 300 mL of water and extracted withethyl acetate (3×60 mL). Organic phases were combined and washed twicewith water, and once with brine, then dried over anhydrous magnesiumsulfate. Rotary evaporation of solvent yielded crude product, which waspurified by silica gel column chromatography (acetic ether:petroleumether=1:1) to yield 1.1 g of pure title product. ¹H NMR (CDCl₃, ppm) δ7.88-7.90 (m, 2H), 7.77-7.79 (m, 2H), 5.06 (dd, 1H, J=5.6, 12.4 Hz),4.02-4.12 (m, 2H), 3.76-3.80 (m, 2H), 2.94-3.02 (m, 1H), 2.72-2.90 (m,2H), 2.29-2.31 (m, 1H), 2.14-2.23 (m, 1H).

Example 22-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(tert-butoxycarbonylamino)-3-methylbutanoate

(S)-2-Boc-amino-3-methyl butyric acid (1.03 g),2-(1-(2-Hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione(1.51 g), and DMAP (20 mg) were dissolved in DCM (30 mL). The mixturewas magnetically stirred at room temperature. DCC (1.10 g) was added inone portion and the mixture was reacted overnight. The mixture wasfiltered to remove dicyclohexylurea. The filter cake washed severaltimes with DCM. The combined filtrates were washed three times withsaturated sodium bicarbonate solution and once with brine (30 mL), driedover anhydrous magnesium sulfate and filtrated. The solvent was removedby rotary evaporation in vacuo. A white solid (430 mg) was obtainedafter silica gel column purification (50% acetic ether/petroleum ether).

Example 32-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-methylbutanoate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(tert-butoxycarbonylamino)-3-methylbutanoate (410 mg) wasdissolved in 30% TFA/DCM (5 mL), the mixture was stirred on a magneticstirrer for 4 hrs at room temperature. White foam was obtained afterreduced pressure distillation to distill off the solvent. The foam wasdissolved in DCM (30 mL). The resultant solution washed with aqueoussaturated sodium bicarbonate solution and brine (30 mL), dried overanhydrous magnesium sulfate, and filtered. Title compound was obtainedas a white solid (260 mg) after rotary evaporation. ¹H NMR (CDCl₃, ppm)δ 7.87-7.91 (m, 2H), 7.76-7.79 (m, 2H), 4.97-5.08 (m, 1H), 4.38-4.43 (m,1H), 4.05-4.30 (m, 3H), 3.25 (dd, 1H, J=4.8, 13.2 Hz), 2.95-3.05 (m,1H), 2.80-2.95 (m, 2H), 2.10-2.20 (m, 1H), 1.90-2.10 (m, 2H), 1.00-1.20(m, 1H), 0.95-0.98 (m, 3H), 0.87-0.91 (m, 3H). MS (m/e): 402 (M+H⁺).

Example 4 2,5-Dioxopyrrolidinyl bromoacetate

Bromoacetic acid (4.30 g) and N-hydroxysuccinimide (4.03 g) weredissolved in DCM (25 ml). The mixture was stirred on a magnetic stirrerat room temperature. DCC was added (7.42 g) in one portion and themixture was reacted overnight. The reaction mixture was filtered toremove dicyclohexylurea. The filter cake washed several times with DCM.The combined filtrates were washed three times with saturated aqueoussodium chloride solution (30 mL/each wash), dried over anhydrousmagnesium sulfate, and filtered. The title compound was obtained as awhite solid (5 g) after rotary evaporation in vacuo.

Example 52-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(2-bromoacetamido)-3-methylbutanoate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-methylbutanoate (1.8 g) was dissolved in DCM (20 mL).2,5-Dioxopyrrolidinyl bromoacetate (1.04 g) was added to the mixture.The mixture was stirred with a magnetic stirrer at room temperature, andreacted overnight. The solvent was stripped in vacuo. White solid (1.3g) was obtained after purification of the crude product on silica gelcolumn (eluted with acetic ether:petroleum ether=1:1).

Example 62-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(2-diethylaminoacetamido)-3-methylbutanoate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(2-bromoacetamido)-3-methylbutanoate (120 mg) was dissolved in DCM(8 mL). The mixture was stirred and diethylamine solution was addedslowly dropwise (0.04 mL). The reaction mixture was stirred additionallyfor 2 hrs at room temperature. The solvent and the residual diethylaminewere removed by rotary evaporation in vacuo. 101 mg of white solid wereobtained after purification of the crude product by silica gel columnchromatography (eluted with acetic ether:petroleum ether=3:1). ¹H NMR(CDCl₃, ppm) δ 7.94 (d, 1H, J=8.4 Hz), 7.87-7.91 (m, 2H), 7.76-7.79 (m,2H), 4.97-5.08 (m, 1H), 4.38-4.43 (m, 1H), 4.05-4.30 (m, 3H), 3.25 (dd,1H, J=4.8, 13.2 Hz), 3.05 (s, 2H), 2.95-3.05 (m, 1H), 2.80-2.95 (m, 2H),2.45-2.58 (m, 4H), 2.10-2.20 (m, 1H), 1.90-2.10 (m, 2H), 1.00-1.20 (m,7H), 0.95-0.98 (m, 3H), 0.87-0.91 (m, 3H).

Example 72-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(2-diethylaminoacetamido)-3-methylbutanoate hydrochloride

The compound (76 mg) obtained from example 6 was dissolved in DCM (10mL) and 15% HCl/methanol solution (5 mL) were added dropwise. Thesolvent was eliminated by rotary evaporation in vacuo and then whitesolid (82 mg) was obtained.

Example 82-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dionebromoacetate

Bromoacetic acid (138.95 mg) and2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (288mg) were dissolved in DCM (20 mL). The mixture was stirred on a magneticstirrer at room temperature. DCC (206 mg) was added in one portion. Themixture was allowed to react overnight. Then, the mixture was filteredto remove dicyclohexylurea. The filter cake washed several times withDCM. The filtrates were combined and then washed three times with brine(30 mL/wash), dried over anhydrous magnesium sulfate, and filtered. Thesolvent was eliminated by rotary evaporation in vacuo and then whitesolid (390 mg) was obtained. ¹H NMR (CDCl₃, ppm) δ 7.88-7.90 (m, 2H),7.77-7.79 (m, 2H), 4.96-5.08 (m, 1H), 4.85 (s, 2H), 4.02-4.12 (m, 2H),3.76-3.80 (m, 2H), 2.94-3.02 (m, 1H), 2.72-2.90 (m, 2H), 2.14-2.23 (m,1H).

Example 92-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione2-(diethylamino)acetate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dionebromoacetate (409 mg) was dissolved in DMF (10 mL). Potassium carbonatepowder (800 mg) was added. Diethylamine solution (0.4 mL) was addeddropwise while stirring. The reaction mixture was stirred for 24 hrs atroom temperature. The solvent and the residual diethylamine were removedby rotary evaporation in vacuo. The resultant solid mixture wassubjected to silica gel column chromatography (eluted with aceticether:petroleum ether=2:1) and then a white solid (128 mg) was obtained.¹H NMR (CDCl₃, ppm) δ 7.87-7.90 (m, 2H), 7.76-7.79 (m, 2H), 4.97-5.04(m, 1H), 4.28-4.33 (m, 2H), 4.08-4.16 (m, 2H), 3.30 (s, 2H), 2.97-3.02(m, 1H), 2.76-2.85 (m, 2H), 2.61-2.68 (m, 4H), 2.10-2.14 (m, 1H),1.02-1.06 (m, 6H). MS (m/e): 416 (M+H⁺).

Example 102-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione2-(diethylamino)acetate hydrochloride

The compound obtained from example 9 (76 mg) was dissolved in DCM (10mL), and 15% HCl/methanol solution (10 mL) was added dropwise. Thesolvent was removed by rotary evaporation in vacuo to afford white solid(80 mg). Solubility of this compound in water was higher than 100 mg/mL.

Example 112-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione2-(dimethylamino)acetate

The title compound was obtained by the method described in example 9with dimethylamine instead of diethylamine. ¹H NMR (CDCl₃, ppm) δ7.87-7.90 (m, 2H), 7.76-7.79(m, 2H), 4.97-5.08(m, 1H), 4.28-4.33(m, 2H),4.08-4.16(m, 2H), 3.30(s, 2H), 2.97-3.02(m, 1H), 2.76-2.85(m, 2H),2.31(s, 6H), 2.10-2.14(m, 1H).

Example 122-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione2-(dimethylamino)acetate hydrochloride

The title compound was obtained by the method of described in exampleten with the title compound of example 11 as a starting material. Thesolubility of the title compound of this example was higher than 100mg/mL in water.

Example 132-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione2-(1-piperidyl)acetate

The title compound was obtained by the method described in example 9with piperidine in place of diethylamine. ¹H NMR (CDCl₃, ppm) δ7.87-7.90 (m, 2H), 7.76-7.79 (m, 2H), 4.97-5.08 (m, 1H), 4.28-4.33 (m,2H), 4.08-4.16 (m, 2H), 3.30 (s, 2H), 2.97-3.02 (m, 1H), 2.76-2.85 (m,2H), 2.31 (s, 6H), 2.10-2.14 (m, 1H).

Example 142-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione2-(1-piperidyl)acetate hydrochloride

The title compound was obtained by the method described in example 10with the title compound of example 13 as starting material. Thesolubility of the title compound of this example in water was more than100 mg/mL.

Example 152-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(diethylamino)-3-methylbutanoate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-methylbutanoate (92 mg) was dissolved in acetonitrile (18mL). Ethyl iodide (74 mg) was added. The mixture was stirred at 80° C.and allowed to react overnight. The solvent was removed by rotaryevaporation in vacuo. White solid (30 mg) was obtained after the crudeproduct was purified by silica gel column (eluted with aceticether:petroleum ether=1:1). ¹H NMR (CDCl₃, ppm) δ 7.86-7.90 (m, 2H),7.76-7.79 (m, 2H), 4.97-5.08 (m, 1H), 4.38-4.43 (m, 1H), 4.05-4.30 (m,3H), 3.15-3.25 (m, 1H), 2.95-3.05 (m, 1H), 2.80-2.95 (m, 2H), 2.45-2.58(m, 4H), 2.10-2.20 (m, 1H), 1.90-2.10 (m, 2H), 1.00-1.20 (m, 7H),0.95-0.98 (m, 3H), 0.87-0.91 (m, 3H).

Example 162-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dioneN-tert-butoxycarbonyl-(S)-2-pyrrolidinecarboxylate

(S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid (374 mg) and2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (500mg) were dissolved in DCM (30 mL). The mixture was stirred on a magneticstirrer at room temperature. DCC (350 mg) and DMAP (25 mg) were added inone portion and the mixture was allowed to react overnight. The mixturewas then filtered to remove dicyclohexylurea. The filter cake washedseveral times with DCM, and the filtrates were combined. The filtrateswere dried over anhydrous magnesium sulfate and filtered. The solventwas removed by rotary evaporation in vacuo. White solid (658 mg) wasobtained after the crude product was purified by silica gel columnchromatography (eluted with chloroform:acetone 9:2).

Example 172-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-pyrrolidinecarboxylate

The compound obtained in example 16 (658 mg) was dissolved in 25%TFA/DCM solution (10 mL). The mixture was stirred on a magnetic stirrerfor 4 hrs at room temperature. DCM and most of TFA were removed byrotary evaporation in vacuo. The foam obtained was dissolved in DCM (50mL). The resultant mixture was washed with saturated sodium bicarbonatesolution and brine, dried over anhydrous magnesium sulfate, andfiltered. The solid obtained (380 mg) was dried in vacuo. ¹H NMR (CDCl₃,ppm) δ 8.0-8.1 (m, 2H), 7.90-8.00 (m, 2H), 5.20-5.28 (m, 1H), 4.59-4.62(m, 1H), 4.30-4.55 (m, 2H), 4.00-4.30 (m, 2H), 3.70-3.85 (m,1H),3.40-3.65 (m, 2H), 2.90-3.12 (m, 2H), 2.70-2.90 (m, 1H), 2.30-2.50 (m,1H), 2.00-2.20 (m, 4H)

Example 182-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(nicotinamido)-3-methylbutanoate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-methylbutanoate (200 mg) and 2,5-dioxopyrrolidinylnicotinate (120 mg) were dissolved in DCM (20 mL). The mixture wasstirred on a magnetic stirrer at room temperature. Triethylamine (1 mL)was added in one portion and the mixture was allowed to react overnight.The reaction mixture was then poured into DCM (30 mL), washed withsaturated sodium bicarbonate solution three times (30 mL/wash) and brine(30 mL), dried with anhydrous magnesium sulfate, and filtered. Thesolvent was removed by rotary evaporation in vacuo. The crude productwas purified by silica gel column chromatography (eluted withchloroform:acetone=5:2) to yield pure title compound (239 mg). ¹HNMR(CDCl₃, ppm) δ 9.04 (d, 1H, J=11.2 Hz), 8.72 (s, 1H), 8.13 (d, 1H, J=8.0Hz), 7.87-7.90 (m, 2H), 7.76-7.78 (m, 2H), 7.41 (dd, 1H, J=8.0, 11.2Hz), 6.73 (d, 1H, J=9.6 Hz), 5.86-5.98 (m, 2H), 5.05-5.08 (m, 1H),3.00-3.15 (m, 1H), 2.80-2.95 (m, 2H), 2.12-2.28 (m, 1H), 2.10-2.20 (m,2H), 0.97-1.05 (m, 3H), 0.85-0.88 (m, 3H).

Example 192-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-phenylpropanoate

1) 2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(tert-butoxycarbonylamino)-3-phenylpropanoate

2-(S)-Boc-amino-3-phenylpropanoic acid (265 mg) and2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (302mg) were dissolved in DCM (50 mL). The mixture was stirred on a magneticstirrer at room temperature. DCC (227 mg) and DMAP (20 mg) were added inone portion and the mixture was allowed to react overnight.Dicyclohexylurea was filtered off. The filter cake washed with DCMseveral times. The filtrates were combined, washed with brine threetimes (30 mL/wash), dried over anhydrous magnesium sulfate, andfiltered. The solvent is eliminated by rotary evaporation in vacuo. Thecrude product was purified by silica gel column chromatography (elutedwith dichloromethane:acetone=5:2) to yield 522 mg of pure titlecompound.

2) 2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-phenylpropanoate

2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-(tert-butoxycarbonylamino)-3-phenylpropanoate (100 mg) wasdissolved in 25% TFA/DCM (10 mL) and stirred on a magnetic strirrer atroom temperature. The mixture was allowed to react for 4 hrs. DCM andmost of TFA were removed by rotary evaporation in vacuo. The foamobtained was dissolved in DCM (50 mL). The mixture washed with saturatedsodium bicarbonate and brine, dried over anhydrous magnesium sulfate,and filtered. 52 mg of a solid was obtained after drying in vacuo. ¹HNMR (CDCl₃, ppm) δ 7.80-7.90 (m, 2H), 7.70-7.80 (m, 2H), 7.10-7.35 (m,5H), 4.95-5.12 (m, 1H), 4.35-4.45 (m, 1H), 4.15-4.25 (m, 2H), 4.00-4.15(m, 1H), 3.65-3.72 (m, 1H), 2.95-3.10 (m, 2H), 2.75-2.90 (m, 3H),2.12-2.20 (m, 1H).

Example 202-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dionenicotinate

The title compound was obtained by the method described in example 19with nicotinic acid instead of(S)-2-(tert-butoxycarbonylamino)-3-phenylpropanoate. ¹H NMR (CDCl₃, ppm)δ 9.2 (s, 1H), 8.78 (d, 1H, J=4.0 Hz), 8.29 (d, 1H, J=8.0 Hz), 7.87-7.90(m, 2H), 7.75-7.78 (m, 2H), 7.41 (dd, 1H, J=4.0, 8.0 Hz), 4.97-5.08 (m,1H), 4.28-4.33 (m, 2H), 4.08-4.16 (m, 2H), 3.30 (s, 2H), 2.97-3.02 (m,1H), 2.76-2.85 (m, 2H), 2.31 (s, 6H), 2.10-2.14 (m, 1H).

Example 212-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dioneN-ethyl-(S)-2-pyrrolidinecarboxylate

The title compound was obtained by the preparation method described inexample 15 with2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-pyrrolidinecarboxylate instead of2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione(S)-2-amino-3-methylbutanoate.

Example 222-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

The title compound was obtained by the preparation method described inexample 1 by the reaction of2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and 4-chlorobutanol. ¹HNMR (CDCl₃, ppm) δ 7.88-7.91 (m, 2H), 7.76-7.78 (m, 2H), 4.95-5.05 (m,1H), 3.82-3.88 (m, 2H), 3.53-3.60 (m, 2H), 2.94-3.02 (m, 1H), 2.72-2.86(m, 2H), 2.9-2.20 (m, 1H), 1.64-1.88 (m, 4H).

Example 232-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dionenicotinate

The title compound was obtained by the method described in example 19 ina reaction of nicotinic acid and2-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione. MS(m/e): 436 (M+H).

Example 242-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione

The title compound was obtained by the method described in example 1 ina reaction of 4-nitro-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dioneand chloroethanol. MS (m/e): 347.

Example 252-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione

The title compound was obtained by the method described in example 1 ina reaction of 4-nitro-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dioneand 2-methoxyethyl 4-methylbenzenesulfonate. MS (m/e): 361.

Example 264-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

10% Pd/C (30 mg) was added to a THF solution of2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione(150 mg). The reaction vessel was pressurized with hydrogen gas at 5times the atmospheric pressure. After 6 hrs of reaction time, thecatalyst was filtered off and solvent was removed by rotary evaporationin vacuo yielding 138 mg of light-yellow solid. MS (m/e): 317.

Example 274-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

10% Pd/C (50 mg) was added to a THF solution of2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione(260 mg). The reaction vessel was pressurized with hydrogen gas at 5times the atmospheric pressure. After 6 hrs of reaction time, thecatalyst was filtered off and solvent was removed by rotary evaporationin vacuo yielding 218 mg of light-yellow solid. MS (m/e): 332 (M+H⁺).

Example 28N-(2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-7-yl)acetamide

Acetic anhydride (0.5 mL) and DMAP (3 mg) were added to a THF solutionof4-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione(50 mg). The reaction mixture was allowed to react for 8 hrs at roomtemperature. Then, DCM (15 mL) were added and solvents were removed byrotary evaporation in vacuo. The residue was washed with 0.5 N aqueoushydrochloric acid, saturated sodium bicarbonate, and brine, dried overanhydrous magnesium sulfate, and filtered. Pale white solid (38 mg) wasobtained after removal of solvent by rotary evaporation in vacuo. MS(m/e): 374 (M+H).

Example 294-(dimethylamino)-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

Methyl iodide (0.1 mL) and potassium carbonate powder (300 mg) wereadded to a DMF solution of4-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione(50 mg). The reaction mixture was allowed to stir for 48 hrs at roomtemperature. Then, 30 mL of water was added in for dilution. The mixturewas extracted with ethyl acetate (3×20 mL). The organic phases werecombined, washed twice with water and once with brine, dried overanhydrous magnesium sulfate and filtered. A crude product was obtainedby rotary evaporation in vacuo and was purified further by silica gelcolumn chromatography (eluted with acetic ether eluate:petroleumether=2:1) to yield 32 mg of pure product. MS (m/e): 360 (M+H⁺).

Example 304-fluoro-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione

The title compound was obtained by the method described in example 1 inthe reaction of4-fluoro-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and2-methoxyethyl 4-methylbenzenesulfonate. MS (m/e). MS (m/e): 333.

Example 314,5,6,7-tetrafluoro-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione

The title compound was obtained by the method described in example 1 bythe reaction of4,5,6,7-tetrafluoro-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione and2-methoxyethyl 4-methylbenzenesulfonate. MS (m/e): 387.

Example 322-(1-(4-(4-methylbenzene)sulfonyloxybutyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione

2-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione wasdissolved in pyridine to which p-toluensulfonyl chloride was previouslyadded. The mixture was reacted at 50° C. for 18 hours. The solvent waseliminated by rotary evaporation in vacuo. Then, 30 mL of saturatedsodium bicarbonate solution were added to the residue. The resultantmixture was extracted with ethyl acetate (3×20 mL). Organic phases werecombined, washed twice with water and once with brine, dried overanhydrous magnesium sulfate, and filtered. Crude product obtained afterrotary evaporation in vacuo was used directly in the next example.

Example 332-(1-(4-(diethylamino)butyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

The title compound was obtained by the method described in example 9 inthe reaction of2-(1-(4-(4-methylbenzene)sulfonyloxybutyl)-2,6-dioxopiperidin-3-yl)isoindolin-1,3-dione,and diethylamine. MS (m/e): 386(M+H).

What is claimed is:
 1. A compound of the formula (I), or an organic orinorganic salt thereof,

wherein n is 1, 2, 3, 4, 5 or 6, R¹ represents from one to four of thesame or different substituents selected from F, Cl, Br, C₁₋₄ alkyl, OH,OC₁₋₄ alkyl, NO₂, NHC(O)C₁₋₄ alkyl, NH₂, NH(C₁₋₄ alkyl), N(C₁₋₄ alkyl)₂;R² represents OR³, NR³R⁴, or N(R³)COR⁴; and R³ and R⁴ representindependently and at each occurrence H or C₁₋₄ alkyl.
 2. The compound ofclaim 1, wherein n is 1, 2, or
 3. 3. The compound of claim 1, wherein R¹represents from one to four of the same or different substituentsselected from F, NO₂, NH₂, NHCH₃, NHCH₂CH₃, N(CH₃)₂, N(CH₂CH₃)₂,—NHCOCH₃.
 4. The compound of claim 1, wherein R² represents OH, OCH₃,OCH₂CH₃, OCH₂CH₂CH₃, OCH(CH₃)₂, NH₂, NHCH₃, NHCH₂CH₃, N(_(CH) ₃)₂, orN(CH₂CH₃)₂.
 5. A compound of claim 1, wherein the compound is4-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.6. A compound of claim 1, wherein the compound is4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.7. A compound of claim 1, wherein the compound is4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione ;or2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione.8. A compound of claim 1, wherein the compound is5-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;4-(dimethylamino)-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-fluoro-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;or4,5,6,7-tetrafluoro-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione.9. A compound of claim 1, wherein the compound is2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-5-nitroisoindoline-1,3-dione;4,5,6,7-tetrafluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;or4-(dimethylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.10. A compound of claim 1, wherein the compound is2-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione. 11.A compound of claim 1, wherein the compound is4-amino-2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;or4-(dimethylamino)-2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.12. A compound of claim 1, wherein the compound is4-amino-2-(2,6-dioxo-1-(2-propoxyethyl)piperidin-3-yl)isoindoline-1,3-dione;or4-(methylamino)-2-(2,6-dioxo-1-(2-propoxyethyl)piperidin-3-yl)isoindoline-1,3-dione.13. A compound of claim 1, wherein the compound is4-(dimethylamino)-2-(1-(3-hydroxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;or4-amino-2-(1-(3-hydroxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.14. A compound of claim 1, wherein the compound is4-(dimethylamino)-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-amino-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;or5-amino-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.15. A compound of claim 1, wherein the compound is2-(1-(4-(diethylamino)butyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.16. A compound of claim 1, wherein the compound is4-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-fluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;5-amino-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;or2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;5-amino-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;4-(dimethylamino)-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-fluoro-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;or4,5,6,7-tetrafluoro-2-(1-(2-methoxyethyl)-2,6-dioxopiperidin-3-yl)-4-nitroisoindoline-1,3-dione;2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-5-nitroisoindoline-1,3-dione;4,5,6,7-tetrafluoro-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;or4-(dimethylamino)-2-(1-(2-hydroxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(4-hydroxybutyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-amino-2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)-4-(methylamino)isoindoline-1,3-dione;or4-(dimethylamino)-2-(1-(2-ethoxyethyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-amino-2-(2,6-dioxo-1-(2-propoxyethyl)piperidin-3-yl)isoindoline-1,3-dione;or4-(methylamino)-2-(2,6-dioxo-1-(2-propoxyethyl)piperidin-3-yl)isoindoline-1,3-dione;4-(dimethylamino)-2-(1-(3-hydroxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;or4-amino-2-(1-(3-hydroxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-(dimethylamino)-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;4-amino-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;or5-amino-2-(1-(3-methoxypropyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione;2-(1-(4-(diethylamino)butyl)-2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione.17. A method for preparation of a compound of claim 1 comprisingreacting a compound of formula (VI)

with a compound of formula Z—CH₂(CH₂)_(n)—R² in the presence of a base,wherein the numerical values, atoms and groups represented by n, R¹, andR² are the same as those in claim 1, and Z represents Br, or Cl.